Small-interference RNA gene knockdown of pancreatitis-associated proteins in rat acute pancreatitis

Yin Yao Lin, Domenico Viterbo, Cathy M. Mueller, Albert E. Stanek, Tamar Smith-Norowitz, Hazel Drew, Raj Wadgaonkar, Michael E. Zenilman, Martin H. Bluth

Research output: Contribution to journalArticlepeer-review

Abstract

OBJECTIVES: Pancreatitis-associated proteins (PAPs) are induced in acute pancreatitis and antisense-mediated gene knockdown of PAP decreased PAP gene expression and worsened pancreatitis. Here, we investigated the effect of a more stable inhibition of PAP using small-interference RNA gene knockdown in vitro and in an in vivo model of experimental pancreatitis. METHODS: Pancreatitis-associated protein-specific siRNA was administered to AR42J cell cultures or rats induced with pancreatitis. Controls included administration of scrambled siRNA or vehicle alone. After 24 hours, cells and pancreata were harvested and assessed for PAP (PAP 1, PAP 2, PAP 3) gene expression and pancreatitis severity. RESULTS: In vitro, PAP protein, and mRNA levels were reduced (PAP 1, 76%; PAP 2, 8%; PAP 3, 24%) in cells treated with PAP siRNA. In vivo, PAP 1, and PAP 3 expressions were reduced (PAP 1, 36%; PAP 3, 66%) in siRNA-treated rats; there was no difference in PAP 2 isoform mRNA expression and serum protein levels. Serum amylase and lipase levels decreased (≥50%) after administration of siRNA; interleukin (IL) 1β, IL-4, and IL-6 increased, whereas C-reactive protein and tumor necrosis factor-α decreased when compared with vehicle control. Administration of PAP siRNA correlated with worsening histopathology. CONCLUSIONS: siRNA-mediated gene knockdown of PAP worsens pancreatitis. Differences in gene knockdown technology may provide different approaches to study gene function.

Original languageEnglish (US)
Pages (from-to)402-410
Number of pages9
JournalPancreas
Volume36
Issue number4
DOIs
StatePublished - May 2008

Keywords

  • AR42J
  • Antisense
  • Cytokines
  • Gene knockdown
  • In vivo
  • Pancreatitis
  • Pancreatitis-associated proteins
  • Severity
  • Sodium taurocholate
  • siRNA

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism
  • Hepatology
  • Endocrinology

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